Comparison of Cytomegalovirus Viral Load Measure by Real-Time PCR With pp65 Antigenemia for the Diagnosis of Cytomegalovirus Disease in Solid Organ Transplant Patients

Hernando, Susana; Folgueira, Lola; Lumbreras, Carlos; Juan, Rafael San; Maldonado, Sonia; Prieto, Columbiana; Babiano, M.J.; Delgado, Juan Marcelo; Andrés, Amado Philip de; Moreno, Enrique; Aguado, José María; Otero, Joaquín R.

doi:10.1016/j.transproceed.2005.10.087

Cited by 28 publications

(17 citation statements)

References 4 publications

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“…Quantitative PCR methods are now routinely used for determining HCMV loads, and monitoring of viral DNAemia certainly has an important role in the clinical management of transplant patients. However, active HCMV disease does not always correlate with viral load detection, and a proportion of patients exhibit a detectable viral load without developing symptomatic clinical disease and are thus unnecessarily preemptively treated with toxic antiviral mediations (105,110). Antigen-specific CD8 ϩ T cells are clearly crucial components of the immune response against HCMV.…”

Section: Virus-specific T Cells For Monitoring Of Hcmv Infectionmentioning

confidence: 99%

Immunobiology of Human Cytomegalovirus: from Bench to Bedside

2009

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SUMMARY Following primary infection, human cytomegalovirus (HCMV) establishes lifelong latency and periodically reactivates without causing symptoms in healthy individuals. In the absence of an adequate host-derived immune response, this fine balance of permitting viral reactivation without causing pathogenesis is disrupted, and HCMV can subsequently cause invasive disease and an array of damaging indirect immunological effects. Over the last decade, our knowledge of the immune response to HCMV infection in healthy virus carriers and diseased individuals has allowed us to translate these findings to develop better diagnostic tools and therapeutic strategies. The application of these emerging technologies in the clinical setting is likely to provide opportunities for better management of patients with HCMV-associated diseases.

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Section: Virus-specific T Cells For Monitoring Of Hcmv Infectionmentioning

confidence: 99%

Immunobiology of Human Cytomegalovirus: from Bench to Bedside

2009

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“…CMV infection is the most frequent cause of infection in patients who have undergone solid-organ transplantation; in varying series, its incidence ranges from 25% to 85% [1,6]. Home-brew (i.e., in-house developed tests) molecular assays are unique for every laboratory.…”

Section: CMVmentioning

confidence: 99%

“…BKV and EBV are not detected in cell cultures by routine diagnostic virology methods. Detection of CMV in blood specimens can be qualitatively recognized rapidly (16 h after inoculation) in shell vial cultures, but quantitation of the CMV load with this technology is cumbersome and impractical.The first laboratory method for quantitation of CMV load was the antigenemia test [4] jective interpretation of the test results [5][6][7]. Importantly, both shell vial cell cultures (qualitative) and antigenemia tests (quantitative) have been shown in comparative studies to be less sensitive than PCR.…”

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confidence: 99%

“…Importantly, both shell vial cell cultures (qualitative) and antigenemia tests (quantitative) have been shown in comparative studies to be less sensitive than PCR. In addition, DNAemia was detected in an earlier stage of nucleic acid amplification [6][7][8][9][10].As an alternative to cell culture and the antigenemia test, implementation of molecular techniques for the rapid and sensitive detection of nucleic acid targets has yielded new levels of capabilities for laboratory diagnosis of many microbial infections [11]. Implementation of these tests has been facilitated by the availability of real-time PCR instrumentation with automation of nucleic acid-target amplification and amplicondetection steps in a "closed system" (i.e., reaction tubes never opened during or after amplification).…”

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confidence: 99%

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Quantitative Real-Time Polymerase Chain Reaction for Evaluating DNAemia due to Cytomegalovirus, Epstein-Barr Virus, and BK Virus in Solid-Organ Transplant Recipients

Smith¹,

Espy²,

Mandrekar³

et al. 2007

Clinical Infectious Diseases

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Testing for cytomegalovirus-, Epstein-Barr virus-, and BK virus-specific gene targets in specimens from solid-organ transplant recipients for DNA by quantitative real-time polymerase chain reaction has been implemented in many diagnostic facilities. This technology provides rapid, accurate, and reproducible results for early detection, monitoring, and medical management of patients with these infections. Because these assays are becoming commonly used in clinical practice, the technical variables associated with specimen processing (e.g., nucleic acid extraction, gene target, and result reporting), amplification, and unique patient characteristics (e.g., age, sex, underlying diseases, immune status, and immunosuppressive regimens received) are factors that may influence the understanding and interpretation of test results. We emphasize the need for standardization of existing variables through parallel comparative and proficiency testing, uniform units for expressing results, to provide for clinical correlation with the results of these molecular assays.Cytomegalovirus (CMV), Epstein-Barr virus (EBV), and BK virus (BKV) are common pathogens and major causes of morbidity and mortality in patients who have received a solid-organ transplant [1][2][3]. Infection with these viruses is indicated by demonstrating the presence of the virus in tissue and/or blood specimens. Because all of these viruses (i.e., CMV, EBV, and BKV) contain DNA, the term "DNAemia" certainly applies. The more general "viremia" can apply to a bloodstream infection caused by a DNA-or RNA-containing virus. BKV and EBV are not detected in cell cultures by routine diagnostic virology methods. Detection of CMV in blood specimens can be qualitatively recognized rapidly (16 h after inoculation) in shell vial cultures, but quantitation of the CMV load with this technology is cumbersome and impractical.The first laboratory method for quantitation of CMV load was the antigenemia test [4] jective interpretation of the test results [5][6][7]. Importantly, both shell vial cell cultures (qualitative) and antigenemia tests (quantitative) have been shown in comparative studies to be less sensitive than PCR. In addition, DNAemia was detected in an earlier stage of nucleic acid amplification [6][7][8][9][10].As an alternative to cell culture and the antigenemia test, implementation of molecular techniques for the rapid and sensitive detection of nucleic acid targets has yielded new levels of capabilities for laboratory diagnosis of many microbial infections [11]. Implementation of these tests has been facilitated by the availability of real-time PCR instrumentation with automation of nucleic acid-target amplification and amplicondetection steps in a "closed system" (i.e., reaction tubes never opened during or after amplification). Therefore, for both qualitative and quantitative detection of viruses, automated realtime PCR has generally replaced the time-consuming and laborintensive conventional amplification and detection of products by gel electrophoresis and ...

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“…While many of these rely on fluorescence resonance energy transfer (FRET) to produce real-time detection, 6 several primer-probe chemistries have been used. 5 Various real-time chemistries have been used to target CMV, 4,[7][8][9] including TaqMan, dual hybridization probes, and labeled primer systems. All of these FRET systems rely on either the production or quenching of fluorescence signal; all but the labeled primer chemistries require an initial PCR step with unlabeled primers followed by a separate probe hybridization step to produce signal.…”

mentioning

confidence: 99%